LOCUS       sequence-13754-e        6687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6687)
  AUTHORS   Moore AT, Rankin KE, von Dassow G, Peris L, Wagenbach M, Ovechkina 
            Y, Andrieux A, Job D, Wordeman L
  TITLE     MCAK associates with the tips of polymerizing microtubules.
  JOURNAL   J Cell Biol. 2005 May 9. 169(3):391-7.
  PUBMED    15883193
REFERENCE   2  (bases 1 to 6687)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol. 2005 May 9. 169(3):391-7."
COMMENT     Sequence Label: pMX156
FEATURES             Location/Qualifiers
     source          1..6687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3475..3596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3512..3531)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3566..3585
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3603..4058)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3740..3761)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3952..3971
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4085..4189
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4191..4548
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4191..4382
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4214..4234)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4399..4534
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4583..5377
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4583..5377
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4637..4656)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5247..5266
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5565..5584)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5609..5656
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5985..6573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6474..6493
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctatggt aacatcttta
     1381 aatgaagata atgaaagtgt aactgttgag tggatagaaa atggagatac gaaaggcaaa
     1441 gagattgact tggagagcat cttttcactt aaccctgacc ttgtacctga tgaagatatt
     1501 gagcccagtc cagaactacc tccaccctcg tcatcctcaa aagttaacaa aattgtaaag
     1561 aaccggcgga ctgtggcagc acgggctgtt aagaatgacc ctcccccgag agataataga
     1621 gtggttggtt cagcacgccc tagtcagctt cctgagcaat cgtcttctgc acaacagaat
     1681 gcacgtagaa aatccaattg tgtgaaagaa gtagaaaaat tgcaagaaaa acgagaaaaa
     1741 aggagattgc aacagcaaga acttagagaa aaaagagccc aggatgttga tgctacaaat
     1801 ccaaattacg aaattatgtg tatgatcaga gacttcagag ggagcttgga ttacagaccc
     1861 ctaacaacag cagatcctat tgatgaacat aggatatgtg tttgtgtaag aaaacgacca
     1921 ctcaataaaa aagaaactca aatgaaagat cttgatgtga tcacaatccc tagtaaagat
     1981 gttgtgatgg tccacgaacc caaacagaaa gtagacttaa caaggtactt agaaaatcag
     2041 acatttcggt ttgattacgc cttcgatgac tcagctccta atgaaatggt ttacaggttt
     2101 actgctaggc cactagtgga aacaatattt gaaaggggca tggctacgtg ctttgcttat
     2161 gggcagactg gaagtgggaa aactcatact atgggtggtg acttttcagg aaagaaccag
     2221 gattgttcta agggaattta tgcattagca gctcgcgatg tctttttaat gctgaagaag
     2281 ccaaactata agaaactaga acttcaagta tatgcaacat tttttgaaat ttatagtgga
     2341 aaggtgtttg acttactaaa taggaaaaca aaattaagag tactagaaga tgggaagcag
     2401 caggtccaag tggtggggct gcaggagcgg gaggtgaagt gtgtggagga cgtgctgaag
     2461 ctcatcgaca taggcaacag ctgcagaaca tctggccaaa catctgcaaa tgcacattca
     2521 tctcggagcc atgcagtgtt ccagatcatt ctaagaagga aggggaaact acatggtaaa
     2581 ttttctctca ttgacctggc tgggaacgaa agaggagctg atacatccag tgcagacagg
     2641 caaaccaggc ttgaaggtgc tgaaattaat aaaagccttt tagcactcaa ggagtgcatc
     2701 cgagccttag gtagaaacaa accccatacg cctttccgag cgagcaaact cactcaggtg
     2761 ctgagagatt ccttcatagg tgaaaattct cgtacctgca tgattgccac aatctctcca
     2821 ggaatggcat cctgtgaaaa tactcttaat acgttaagat atgcaaatag ggtgaaagaa
     2881 ttgactgtaa atccagctgc tgctggcgat gtccatccaa taatgcacca tccaccgagc
     2941 cagattgatg acttagagac acagtggggt gtggggagtt ctcctcagag agatgatcta
     3001 aaacttctct gtgaacaaaa tgaagaagaa gtttctccac agctgttcac tttccatgaa
     3061 gctgtctccc agatggtaga gatggaagag caggttgtgg aagatcatag ggcagtgttc
     3121 caggaatcca ttcgctggat agaagatgaa aaggctctcc tagagatgac tgaagaagta
     3181 gactatgatg tagactcata tgctacacaa cttgaagcta ttcttgagca aaaaatagac
     3241 attttaactg agttacgaga taaagtgaaa tcttttcgcg cagcgctaca agaagaagag
     3301 caagcaagca agcaaattaa ccccaagagg ccccgagccc tctaaggatc caccggatct
     3361 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3421 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3481 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3541 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     3601 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3661 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3721 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3781 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3841 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3901 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3961 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     4021 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     4081 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4141 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     4201 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4261 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4321 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4381 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4441 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4501 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4561 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4621 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4681 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4741 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4801 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4861 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4921 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4981 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5041 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5101 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5161 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5221 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5281 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5341 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5401 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5461 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5521 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5581 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5641 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5701 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5761 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5821 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5881 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5941 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6001 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6061 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6121 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6181 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6241 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6301 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6361 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6421 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6481 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6541 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6601 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6661 gtggataacc gtattaccgc catgcat
//
